Journal: Neuropharmacology

Article Title: Nicotine-induced neuroplasticity counteracts the effect of schizophrenia-linked neuregulin 1 signaling on NMDAR function in the rat hippocampus

doi: 10.1016/j.neuropharm.2016.10.021

Figure Lengend Snippet: NMDAR EPSCs evoked by stimulation of the Schaffer collateral afferent were recorded in CA1 pyramidal cells from naive (A, B) and chronic-nicotine-exposed (C, D) rats. Time course of the effect of intracellular application of pYEEI through a patch pipette on NMDAR EPSCs in the absence (A, C) or presence (B, D) of NRG1β (2 nM) is shown as a percent change in amplitude (mean ± SEM). Amplitudes of NMDA EPSCs during the first 5–10 min and the period from 30 to 35 min were used to calculate a percent change for statistical analysis. In naive pyramidal cells, pYEEI enhances NMDAR EPSCs (A) and NRG1β blocks the effect of pYEEI (B). In chronic-nicotine-exposed pyramidal cells, pYEEI enhances NMDAR EPSCs (C), but NRG1β has no effect on this enhancement (D). In this and the following figures, traces above the graph show representative NMDA EPSCs recorded during the first 5 to 10-min period (left) and 30 to 35-min period (right) after whole-cell configuration was established. Each trace is the average of four NMDAR EPSCs. *p < 0.05, ** p < 0.01

Article Snippet: The peptides [Src (p60 c-Src , Upstate, Charlottesville, VA, USA), Src-family activating phosphopeptide (pYEEI; Invitrogen-Biosource, Camarillo, CA, USA), Src inhibitor peptide (40–58) (custom synthesis), Fyn (Millipore, Dundee, UK and Signalchem, Richmond, Canada), Yes (Signalchem, Richmond, Canada), and Lyn (Signalchem)] were directly applied into the pyramidal cells by diffusional exchange through the patch pipettes and neuregulin 1β (NRG1β) (Shenandoah Biotechnology, Warwick, PA, USA) was bath-applied.

Techniques: Transferring

Journal: Neuropharmacology

Article Title: Nicotine-induced neuroplasticity counteracts the effect of schizophrenia-linked neuregulin 1 signaling on NMDAR function in the rat hippocampus

doi: 10.1016/j.neuropharm.2016.10.021

Figure Lengend Snippet: Pharmacologically isolated GluN2B-NMDAR EPSCs (A, C) and GluN2A-NMDAR EPSCs (B) were recorded in CA1 pyramidal cells of naive (A) or chronic-nicotine-exposed (B, C) rats. Time course of the effect of intracellular application of pYEEI (A) or Fyn (B) in the presence of NRG1β (2 nM) on GluN2B-NMDAR EPSCs (A) or GluN2A-NMDAR EPSCs (B) is shown as a percent change in amplitude (mean ± SEM). (C) Time course of the effect of application of NRG1β and AP5 on GluN2B-NMDAR EPSCs in chronic-nicotine-exposed pyramidal cells is shown as a percent change in amplitude (mean ± SEM). Amplitudes of GluN2B-NMDAR EPSCs (A) or GluN2A-NMDAR EPSCs (B) during the first 5–10 min and the period from 30 to 35 min were used to calculate a percent change for statistical analysis. In (C), amplitudes of GluN2B-NMDAR EPSCs during the first −10 – −5 min and the period from 30 to 35 min were used to calculate a percent change for statistical analysis. In naive rats, NRG1β blocks the pYEEI-mediated enhancement of GluN2B-NMDAR EPSCs (A). In chronic-nicotine-exposed rats, NRG1β has no effect on Fyn-mediated enhancement of GluN2A-NMDAR EPSCs (B) and basal GluN2B-NMDAR EPSCs (C), which are completely blocked by the NMDAR antagonist AP5. **p < 0.01

Article Snippet: The peptides [Src (p60 c-Src , Upstate, Charlottesville, VA, USA), Src-family activating phosphopeptide (pYEEI; Invitrogen-Biosource, Camarillo, CA, USA), Src inhibitor peptide (40–58) (custom synthesis), Fyn (Millipore, Dundee, UK and Signalchem, Richmond, Canada), Yes (Signalchem, Richmond, Canada), and Lyn (Signalchem)] were directly applied into the pyramidal cells by diffusional exchange through the patch pipettes and neuregulin 1β (NRG1β) (Shenandoah Biotechnology, Warwick, PA, USA) was bath-applied.

Techniques: Isolation

Journal: bioRxiv

Article Title: Stromal NRG1 in luminal breast cancer defines pro-fibrotic and migratory cancer-associated fibroblasts

doi: 10.1101/2020.04.06.026971

Figure Lengend Snippet: A , expression of NRG1 in breast cancer subtypes (PAM50) extracted from METABRIC and TCGA datasets. Boxes indicate mean +/- quartiles and minimum and maximum values are represented by bars. Only statistically significant comparisons with luminal subtypes are depicted. ANOVA multiple comparison test (*** P < 0.001). B , expression of NRG1 in the epithelial and stromal compartment in 4 laser-capture microdissected (LCM) breast cancer datasets (GSE10797, n = 28; GSE14548, n = 14; GSE35019, n = 53; and GSE83591, n = 39). Boxes indicate mean +/- quartiles and minimum and maximum values in bars. Two-tailed paired Student’s t-test (* P < 0.05; ** P < 0.01; *** P < 0.001). C , viability of T47D (black) and MCF7 (grey) measured 72h after treatment with HER3 mAb lumretuzumab or pertuzumab at indicated doses. Values represent median of 3 independent experiments (n = 5). U-Mann Whitney two-tailed test was applied. No significant differences were observed. D , ectopic NRG-1β (50ng/mL) was added to T47D and MCF7 cell lines and viability quantified. The untreated control was set to 100% (not shown) and used to normalize the results from conditions where cells had been preincubated for 1 hour with lumretuzumab (Lum) or pertuzumab (Pert) at 10µg/mL. Two-tailed unpaired Student’s t-test (* P < 0.05; ** P <0.01, *** P <0.001) comparing each treatment with untreated condition. Bars represent average of two independent experiments +/- s.e.m. E , representative Western blot showing total levels and phosphorylation of HER3 and downstream effectors AKT and ERK1/2, 5min after addition of NRG-1β (50ng/ml). Some samples were either pre-incubated with mAbs lumretuzumab (Lum) or pertuzumab (Pert) at 10µg/ml for one hour.

Article Snippet: Prior to addition of CAF-CM or human recombinant NRG-1β (4711, BioCat), cells were pre-treated with either lumretuzumab or pertuzumab (10μg/ml) for 30min in low serum media (1% FCS).

Techniques: Expressing, Two Tailed Test, MANN-WHITNEY, Western Blot, Incubation

Journal: bioRxiv

Article Title: Stromal NRG1 in luminal breast cancer defines pro-fibrotic and migratory cancer-associated fibroblasts

doi: 10.1101/2020.04.06.026971

Figure Lengend Snippet: A , heatmap representing relative phosphorylation values of HER3, AKT and ERK1/2 in T47D and MCF7 cells after addition of CAF conditioned media (CM) for 5min with or without pre-incubation with lumretuzumab (Lum) (10µg/ml). Values represent median of three technical replicates and three biological replicates. Color intensities are ranked per each antibody (red = maximum, blue = minimum). B , relative proliferation of T47D and MCF7 cancer cells after 72h with different CAF-CM with lumretuzumab (red squares) or untreated (black circles). Dots represent mean +/- s.e.m of three independent replicates (n = 4). P values were determined by two-tailed U-Mann Whitney test for each CM (* P < 0.01; ** P < 0.001; *** P < 0.0005; **** P <0.0001). C , percentage of closure in a scratch assay of T47D or MCF7 cancer cells, after 21h of treatment with 10μg/ml lumretuzumab (red) or untreated (black), and with conditioned media (CM) of indicated CAFs. DMEM-F12 1% FCS was used as negative control and NRG-1β (50ng/ml) as positive control. Box plots correspond to the mean and s.e.m. of 2 independent experiments (n = 6 technical replicates). Two-tailed U-Mann Whitney test for each CM (* P < 0.01; **P < 0.001; *** P < 0.0005; **** P <0.0001).

Article Snippet: Prior to addition of CAF-CM or human recombinant NRG-1β (4711, BioCat), cells were pre-treated with either lumretuzumab or pertuzumab (10μg/ml) for 30min in low serum media (1% FCS).

Techniques: Incubation, Two Tailed Test, MANN-WHITNEY, Wound Healing Assay, Negative Control, Positive Control

Journal: Experimental and Therapeutic Medicine

Article Title: Expression and secretion of neuregulin-1 in cardiac microvascular endothelial cells treated with angiogenic factors

doi: 10.3892/etm.2018.5811

Figure Lengend Snippet: NRG-1β secretion in HCMECs in different groups. (A) NRG-1β secretion in HCMECs with angiogenic factors stimulation under normal conditions. (B) NRG-1β secretion in HCMECs with angiogenic factors stimulation under Hypo/SD conditions. *P<0.05 vs. the control group; #P<0.05 vs. the Hypo/SD only group. HCMECs, human cardiac microvascular endothelial cells; NRG-1β, neuregulin-1β; Hypo/SD, hypoxia/serum deprivation.

Article Snippet: Cells and reagents HCMECs were obtained from ScienCell Research Laboratories (Carlsbad, CA, USA); anti-ErbB2 rabbit monoclonal antibody, anti-ErbB3 rabbit monoclonal antibody and anti-ErbB4 rabbit monoclonal antibody were obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA); anti-NRG-1 rabbit polyclonal antibody was obtained from R&D Systems (Minneapolis, MN, USA); anti-GAPDH rabbit polyclonal antibody was obtained from Signalway Antibody LLC (College Park, MD, USA); VEGF and Ang-1 were obtained from ProSpec-Tany TechnoGene Ltd. (Nessziona, Israel); Ang-2 was obtained from Peprotech (Oak Park, CA, USA); NRG-1β ELISA kit was obtained from RayBiotech (Norcoss, GA, USA).

Techniques: